Study of Neuropeptide F Function in Drosophila

果蝇神经肽F功能的研究

• 批准号: 6635314
• 负责人: PING SHEN
• 金额: $ 24.07万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6635314
• 关键词: Drosophilidae; arthropod genetics; behavioral /social science research tag; biological models; developmental genetics; gene targeting; genetic models; genetically modified animals; immunocytochemistry; in situ hybridization; invertebrate hormone; neuropeptide Y; neuropeptide receptor; neuropeptides; neuroregulation; nutrient intake activity; nutrition related tag; receptor expression

DESCRIPTION (applicant's abstract): Recent molecular and genetic studies indicate that a neuropeptide Y (NPY) mediated signaling pathway, which is conserved among animals including worms and humans, is involved in regulating feeding behavior. However, the mechanism for its regulation is not understood. The long term goal of this proposal is to understand the role of Drosophila neuropeptide F(NPF), a human NPY homolog, in modulating feeding behavior, the underlying mechanism(s), and its functional relationship to vertebrate neuropeptide Y family molecules. Drosophila is the only invertebrate model organism where homologs of NPY and its receptors have been identified. Using whole mount in situ RNA hybridization and immunocytochemistry, we showed that dNPF is expressed in both CNS and gut In larval CNS, dNPFergic neurons of the brain innervate subesophageal, thoracic, and abdominal ganglia, and may thereby act as neurotransmitters for mediating brain control of feeding behavior. We also found that gustatory stimulation by sugars induces dNPF expression in two additional neurons of larval subesophageal ganglion. The expression remains strong long after sugar withdrawl, implicating dNPF in mediating carbohydrate conditioning of fly CNS. The dNPF midgut expression is enhanced by food uptake, and reduced by starvation. One of the putative dNPF receptors is expressed in larval gut, brain and possibly salivary glands. Thus, dNPF may also function as a gut hormone that coordinates feeding associated activities in diverse tissues. In this application, we propose to directly test the specific roles of dNPF in transgenic flies by addressing the following specific aims: 1) Constructing gain-of-function transgenic Drosophila larvae through controlled ectopic dNPF expression and analyzing their feeding behaviors; 2) Cloning and characterization of cis regulatory elements of dNPF using UAStauGFP reporter system; 3) constructing loss-of-function transgenic larvae through targeted functional knockout of dNPF and determining the specific roles of brain, subesophageal ganglion and midgut dNPF in feeding behavior. These studies should provide general insights into the signaling mechanism for feeding regulation in invertebrates as well as higher animals including humans.

描述（申请人的摘要）：最近的分子和遗传学研究 表明神经肽 Y (NPY) 介导的信号通路，即 在包括蠕虫和人类在内的动物中保守，参与调节 摄食行为。然而，其调节机制尚不清楚。 该提案的长期目标是了解果蝇的作用 神经肽 F(NPF) 是一种人类 NPY 同源物，在调节进食行为方面， 潜在机制及其与脊椎动物的功能关系 神经肽Y家族分子。果蝇是唯一的无脊椎动物模型 已鉴定出 NPY 及其受体同源物的生物体。使用 整体原位 RNA 杂交和免疫细胞化学，我们表明 dNPF 在 CNS 和肠道中均表达 在幼虫 CNS 中，dNPFergic 神经元 大脑支配食管下、胸、腹神经节，从而可能 作为调节大脑对进食行为的控制的神​​经递质。我们 还发现糖的味觉刺激会诱导两种细胞中 dNPF 的表达 幼虫食管下神经节的额外神经元。表达式仍然 戒糖后很长一段时间都很强，表明 dNPF 介导碳水化合物 苍蝇中枢神经系统的调节。 dNPF 中肠表达因食物摄取而增强， 并因饥饿而减少。一种推定的 dNPF 受体表达于 幼虫肠道、大脑和可能的唾液腺。因此，dNPF 也可能起到 一种肠道激素，可协调多种喂养相关活动 组织。在这个应用中，我们建议直接测试具体的角色 转基因果蝇中的 dNPF 通过解决以下具体目标：1) 通过控制构建功能获得性转基因果蝇幼虫 异位 dNPF 表达并分析其摄食行为； 2) 克隆和 使用 UAStauGFP 报告基因表征 dNPF 的顺式调控元件 系统; 3）通过靶向构建功能丧失的转基因幼虫 dNPF 的功能性敲除并确定大脑的特定作用， 食管下神经节和中肠 dNPF 在进食行为中的作用。这些研究 应该提供对喂养信号机制的一般见解 无脊椎动物以及包括人类在内的高等动物的调节。